A biosensor for the activity of the "sheddase" TACE (ADAM17) reveals novel and cell type-specific mechanisms of TACE activation.
A biosensor for the activity of the "sheddase" TACE (ADAM17) reveals novel and cell type-specific mechanisms of TACE activation.
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DOI：
10.1126/scisignal.2005680
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发表时间：
2015-02-24
影响因子：
7.3
通讯作者：
Liu X
中科院分区：
文献类型：
--
作者：
Chapnick DA;Bunker E;Liu X
配体脱落作为细胞对不同环境条件作出反应的主要翻译后修饰机制，已受到越来越多的关注。TACEadam17蛋白酶是这种配体脱落的关键介质，调节驱动不同细胞反应的令人印象深刻的细胞外底物的成熟和释放。这种蛋白酶本身是如何被激活的尚不清楚，部分原因是缺乏可用的工具来测量活细胞中具有时间和空间分辨率的TACE活性。我们已经开发了一种基于FRET的TACE活性生物传感器（TSen），能够以高度特异性报告活细胞中的TACE活化动力学。TSen与化学生物学结合使用，以探测各种TACE激活方式对p38和Erk激酶活性的依赖性，以及确定肌动蛋白细胞骨架破坏和TACE激活之间的新联系。这种细胞骨架破坏导致在某些细胞类型中快速和稳健的TACE活化以及TACE在质膜上的积累，从而增加内源性底物的裂解。我们的研究强调了TSen作为了解活细胞中TACE激活机制的工具的多功能性以及肌动蛋白细胞骨架完整性作为TACE活性调节剂的重要性。
Ligand shedding has gained increased attention as a major posttranslational modification mechanism used by cells to respond to diverse environmental conditions. The TACEadam17 protease is a critical mediator of such ligand shedding, regulating the maturation and release of an impressive range of extracellular substrates that drive diverse cellular responses. Exactly how this protease is itself activated remains unclear, in part due to the lack of available tools to measure TACE activity with temporal and spatial resolution in live cells. We have developed a FRET based biosensor for TACE activity (TSen), which is capable of reporting TACE activation kinetics in live cells with a high degree of specificity. TSen was used in combination with chemical biology to probe the dependence of various means of TACE activation on p38 and Erk kinase activities, as well as to identify a novel connection between actin cytoskeletal disruption and TACE activation. Such cytoskeletal disruption leads to rapid and robust TACE activation in some cell types and accumulation of TACE at the plasma membrane, allowing for increased cleavage of endogenous substrates. Our study highlights both the versatility of TSen as a tool to understand the mechanisms of TACE activation in live cells and the importance of actin cytoskeletal integrity as a modulator of TACE activity.
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影响因子：
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